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《中国工程科学》 >> 2014年 第16卷 第9期

大菱鲆神经肽Y基因克隆及其在工厂化养殖饥饿-投喂策略中的表达特征分析

1. 大连海洋大学农业部北方海水增养殖重点实验室,辽宁大连 116023;

2. 中国水产科学研究院黄海水产研究所,农业部海洋渔业可持续发展重点实验室,青岛市海水鱼类种子工程与生物技术重点实验室,山东青岛 266071;

3. 中国科学院动物研究所计划生育生殖生物学国家重点实验室,北京 100101

资助项目 :国家自然科学基金资助项目(30901111);山东省优秀中青年科学家奖励基金项目(BS2011HZ025);现代农业产业技术体系建设专项资金(CARS-50) 收稿日期: 2014-06-30 发布日期: 2014-09-16 08:55:10.000

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摘要

本文通过逆转录聚合酶链式反应(RT-PCR)和末端快速扩增技术(RACE)克隆了大菱鲆神经肽Y(NPY)基因全长为729 bp 的互补脱氧核糖核酸(cDNA)序列,其中5′非翻译区(5′-UTR)为70 bp,3′非翻译区(3′-UTR)为359 bp,开放阅读框(ORF)为300 bp。ORF编码含有99 个氨基酸的蛋白质多肽,N端28 个氨基酸为信号肽,信号肽后是由36 个氨基酸组成的NPY成熟肽。针对大菱鲆NPY蛋白质与其他物种的同源性进行比较,结果显示大菱鲆NPY与其他物种的同源性在53.9 %~98 %,特别是与同属于鲆鲽鱼类的牙鲆和美洲拟鲽的同源性高达97 %~98 %,表现出很高的保守性。采用实时定量PCR对大菱鲆NPY基因的表达特征进行分析,结果表明,大菱鲆NPY基因在神经组织和外围组织中都有表达,其中在下丘脑中的表达量最高。饥饿实验中,随着饥饿的进行,NPY的表达量显著变化(P<0.05),呈现先升高再降低,再显著升高并维持一段高水平表达后,再次下降的趋势,饥饿后再投喂会使NPY的表达量升高,并且饥饿时间越长,再投喂后NPY的表达水平越高,但是再投喂2 h 后表达量还会再次下降至相似的水平,这都预示着NPY是一种摄食刺激因子,并且是一种短期的调控机制。

关键词

大菱鲆 ; NPY ; 饥饿 ; 投喂 ; 差异表达

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参考文献

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